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Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit

A foot-and-mouth disease virus and structural protein technology, which is applied in the field of the foot-and-mouth disease virus structural protein antibody enzyme-linked immunoassay detection kit and the foot-and-mouth disease virus structural protein VP1 antibody enzyme-linked immune detection kit, can solve the problems of international trade blockade, economic loss, restriction and the like , to achieve the effect of high sensitivity, reduced loss and strong specificity

Active Publication Date: 2015-06-03
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, it will trigger the market circulation of animals and animal products in endemic countries or regions, and greatly block and restrict their international trade, causing huge economic losses to animal husbandry production in endemic countries and regions.

Method used

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  • Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit
  • Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit
  • Foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, preparation of FMD virus structural protein antibody enzyme-linked immunoassay kit coating antigen

[0047] In this experiment, aiming at the difference of domestic foot-and-mouth disease epidemic strains, bioinformatics methods were used to accurately analyze multiple subtypes of foot-and-mouth disease virus O-type VP1 structural protein, and suitable peptides were screened out. The 2 peptides designed for the strain sequence, the sequences of which are shown in sequence 1 in the sequence table and sequence 2 in the sequence, are made into a newer and more complete coating antigen with a purity of about 90%, which can adapt to the characteristics of continuous mutation of foot-and-mouth disease virus and improve Antibody positive detection rate. The polypeptide synthesis method can be a conventional method, and the following method is used in the present invention to synthesize the two polypeptides of the present invention as the coating agent of the kit ...

Embodiment 2

[0078] Embodiment 2, the preparation of foot-and-mouth disease virus structural protein antibody ELISA kit

[0079] Foot-and-mouth disease virus structural protein antibody ELISA kit includes:

[0080] (1) 96-well detachable polystyrene enzyme-linked reaction plate coated with foot-and-mouth disease virus antigen; 2×96 wells.

[0081](2) Horseradish peroxidase-labeled rabbit anti-pig IgG polyclonal antibody (purchased from sigma, Cat. No. A5670), 2 vials (12ml each).

[0082] (3) Positive control serum: using the coated antigen artificially synthesized in Example 1 as the immunogen, the hyperimmune serum prepared by immunizing 25-35-day-old specific pathogen-free (SPF) pigs was added with 1000 U / ml streptavidin and 1000U / ml penicillin, sterilized by 0.2μm filter membrane, used as the positive control serum of the kit. 1 tube (1ml).

[0083] (4) Negative control serum: use normal pig serum without FMD antibody, add 1000U / ml streptomycin and 1000U / ml penicillin, pass through ...

Embodiment 3

[0090] Embodiment 3, the coincidence rate test of foot-and-mouth disease virus structural protein antibody enzyme-linked immunoassay kit

[0091] 1. How to use ELISA Kit for Antibody to Foot-and-Mouth Disease Virus Structural Protein

[0092] 1. Equilibration: Take out the kit stored at 4°C and equilibrate to room temperature for later use; mix liquid reagents before use.

[0093] 2. Dosing: Dilute the concentrated washing solution 20 times with distilled water or deionized water to obtain washing buffer;

[0094] 3. Setting: leave 1 blank well, 3 negative control wells, and 2 positive control wells, and the rest are sample wells to be tested.

[0095] 4. Pre-dilution of the sample to be tested: use the sample diluent to dilute the serum of the sample to be tested, the negative control serum, and the positive control serum at a ratio of 1:21; add 100 μl of sample diluent to each well of the dilution plate, and add 5 μl of the waiting Test samples.

[0096] 5. Adding samples...

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Abstract

The invention discloses a foot and mouth disease virus structural protein antibody enzyme-linked immunosorbent assay kit. The kit comprises an enzyme-linked reaction plate coated by foot and mouth disease virus structural protein VP1 antigenic epitope polypeptides, and an enzyme-labeled antibody. The foot and mouth disease virus structural protein VP1 antigenic epitope polypeptides are a polypeptide represented by the sequence 1 in the sequence list and a polypeptide represented by the sequence 2 in the sequence list. According to the kit, the reaction plate coated by chemically synthesized VP1 antigenic peptide, antigen dose is low, sensitivity is high, and specificity is high. With the kit, whether foot and mouth disease virus infection exists can be highly efficiently detected. The kit provided by the invention has good specificity, high sensitivity, high efficiency, and good market prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, the invention relates to an enzyme-linked immunosorbent detection kit for an antibody to the structural protein of foot-and-mouth disease virus, in particular to an enzyme-linked immunosorbent detection kit for an antibody to the structural protein VP1 of foot-and-mouth disease virus. Background technique [0002] Foot-and-Mouth Disease (FMD) is a global epidemic infectious disease that is widely distributed in the world and spread internationally. The International Bureau of Veterinary Medicine (OIE) listed foot-and-mouth disease as the first of Class A infectious diseases in the revised classification of animal infectious diseases in 1984. Foot-and-mouth disease is a highly contagious zoonotic disease of artiodactyls caused by Foot-and-Mouth Disease Virus (FMDV) infection. It is characterized by acute onset, rapid spread, and high infection rate, resulting in a rapid decline in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/09G01N33/68G01N33/569G01N33/543G01N33/545
CPCC07K14/09G01N33/56983G01N2333/09
Inventor 董春娜齐鹏肖进栗利芳王楠巴利民宋芳张蕾周强郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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